The overall goal of this project is to develop immunoassay and immunoaffinity methods for the detection of bryostatin 1 and its analogs in a variety of biological matrices. The bryostatins have exhibited strong antineoplastic activity in preclinical studies and are currently in Phase I clinical trials. Sensitive analytical methods are needed for establishing the proper dosing regimens in these studies and for maximizing yields during the large scale isolation of these scarce natural products. In addition, the availability of bryostatin immunoassays would facilitate basic research efforts to determine the biosynthesis and actual source of these interesting compounds. Our Phase I results demonstrated the technical feasibility of eliciting anti-brytin responses in rabbits and mice immunized with bryostatin-KLH conjugates. Prototype bryostatin immunoassays were developed utilizing polyclonal rabbit antisera. Phase II research will focus on production of monoclonal antibody-based immunoassays and immunoaffinity methods for bryostatins 1, 2, and 3. Procedures will be developed for determining bryostatin concentrations in various biological matrices, including human plasma and B. neritina extracts.